Combination of UV and green light synergistically enhances the attractiveness of light to green stink bugs Nezara spp.

The southern green stink bug Nezara viridula and its congener N. antennata are important agricultural pests worldwide. These species show positive phototaxis and their compound eyes have high sensitivity to UV and green lights. The attractiveness of monochromatic UV, green lights and combined UV and green light to stink bugs was investigated under field conditions. The number of stink bugs caught increased with the number of UV LEDs, but very few bugs were caught using green light, irrespective of the number of LEDs. However, the most stink bugs were caught when both colors were combined. These results indicate that monochromatic green light is less attractive to Nezara bugs, but when mixed with UV light, it synergistically enhances the attractiveness of UV light. This finding contributes to the construction of reliable and highly specific light traps to monitor Nezara bugs. The addition of green light hardly affected the attractiveness of the UV light to other insects, such as Anomala beetles, which are often caught in light traps. We conclude that the spectral composition of light that is attractive to nocturnal insects depends on the species, hence it is possible to make ecologically friendly light traps that are target specific.

Manipulation of Insect Vectors' Host Selection Behavior by Barley Yellow Dwarf Virus Is Dependent on the Host Plant Species and Viral Co-Infection.

Previous studies have shown that vector-borne viruses can manipulate the host selection behavior of insect vectors, yet the tripartite interactions of pathogens, host plants and insect vectors have been documented only in a limited number of pathosystems. Here, we report that the host selection behavior of the insect vector of barley yellow dwarf virus-PAV (BYDV-PAV) and cereal yellow dwarf virus-RPS (CYDV-RPS) is dependent on the host plant species and viral co-infection. This study shows that a model cereal plant, Brachypodium distachyon, is a suitable host plant for examining tripartite interactions with BYDV-PAV and CYDV-RPS. We reveal that BYDV-PAV has a different effect on the host selection behavior of its insect vector depending on the host plant species. Viruliferous aphids significantly prefer non-infected plants to virus-infected wheat plants, whereas viral infection on a novel host plant, B. distachyon, is not implicated in the attraction of either viruliferous or nonviruliferous aphids. Furthermore, our findings show that multiple virus infections of wheat with BYDV-PAV and CYDV-RPS alter the preference of their vector aphid. This result indicates that BYDV-PAV acquisition alters the insect vector's host selection, thereby varying the spread of multiple viruses.

Elucidation of the whole carotenoid biosynthetic pathway of aphids at the gene level and arthropodal food chain involving aphids and the red dragonfly.

BACKGROUND: Aphids can be positioned as robust pest insects in farming and as ones of the model organisms for arthropods in molecular biology. Carotenoids are pigments that protect organisms from photooxidative damage caused by excessive light. Aphids were shown to possess genes of fungal origin for carotenoid biosynthesis, whereas a little knowledge was available about the functions of the genes and the biosynthetic pathway. Even carotenoid species contained in aphids were not enough understood. Main purpose of this study is to clarify these insufficient findings. RESULTS: The whole carotenoid biosynthetic pathway of the pea aphid (Acyrthosiphon pisum) was elucidated at the gene level, through comprehensive functional analysis of its carotenogenic genes, using Escherichia coli that synthesized carotenoid substrates, along with structural and quantitative analysis of carotenoids from various aphid species. Four genes were needed to synthesize all carotenoids accumulated in aphids from geranylgeranyl diphosphate. The tor gene mediated desaturation reaction from phytoene to 3,4-didehydrolycopene. It was revealed that a gene designated ApCrtYB3, which was considered to have functionally evolved in aphids, can convert lycopene into uncommon carotenoids with the γ-ring such as (6'S)-ß,γ-carotene and γ,γ-carotene. We further demonstrated that the atypical carotenoids work as ecological indicators for estimating the food chain from aphids to predatory arthropods, and showed that aphids contributed with significant levels to the food chain from insect herbivores to several predatory arthropods, i.e., the red dragonfly (Sympetrum frequens; adults), seven-spotted ladybird (Coccinella septempunctata), and two spiders, Oxyopes sertatus and Nephila clavata. Gut microflora of the dragonfly (mature adults) was also found to include endosymbiotic bacteria such as Serratia symbiotica specific to the black bean aphid (Aphis fabae). CONCLUSIONS: We revealed the whole carotenoid biosynthetic pathway of aphids, including functional identification of the corresponding genes. Subsequently, we showed that arthropodal food chain can be estimated using the uncommon carotenoids of aphids as ecological indicators. This result indicated that aphids made significant contributions to the food chain of several predatory arthropods including the red-dragonfly adults. Aphids are likely to be positioned as an important "phytochemicals" source for some predatory insects and arachnids, which are often active under bright sunlight.

Maternal hatching synchronization in a subsocial burrower bug mitigates the risk of future sibling cannibalism.

Sibling cannibalism-the killing and consumption of conspecifics within broods-carries a high risk of direct and inclusive fitness loss for parents and offspring. We reported previously that a unique vibrational behavior shown by the mother of the subsocial burrower bug, Adomerus rotundus (Heteroptera: Cydnidae), induced synchronous hatching. Maternal regulation may be one of the most effective mechanisms for preventing or limiting sibling cannibalism. Here, we tested the hypothesis that synchronous hatching induced by maternal vibration in A. rotundus prevents sibling cannibalism. Mothers and their mature egg masses were allocated to three groups: synchronous hatching by maternal vibration (SHmv), synchronous hatching by artificial vibration (SHav), and asynchronous hatching (AH). We then investigated the influence of each hatching strategy on the occurrence of sibling cannibalism of eggs and early-instar nymphs in the laboratory. No difference in the proportion of eggs cannibalized was observed among the three groups. However, the proportion of nymphs cannibalized was higher in the AH group than in the SHmv group. The difference in the number of days to first molting within clutch was significantly higher in the AH group than in the SHmv group. Junior nymphs were sometimes eaten by senior nymphs. However, immediately after molting, senior nymphs were at a high risk of being eaten by junior nymphs. Our results indicate that synchronous hatching of A. rotundus is necessary to mitigate the risk of sibling cannibalism.

Adults of Lasioderma serricorne and Stegobium paniceum (Anobiidae: Coleoptera) Are Attracted to Ultraviolet (UV) Over Blue Light LEDs.

Two species, the cigarette beetle Lasioderma serricorne (F.) and the drugstore beetle Stegobium paniceum (L.), are particularly important stored-product pests because they damage dry food. A previous study showed that L. serricorne adults are attracted more to ultraviolet (UV) and blue light wave ranges more than others such as turquoise, green, yellow, red, and warm white. However, the previous study did not equalize the amounts of light. The study also evaluated the attractiveness by the numbers of L. serricorne individuals that were lured to LED lights in a small box in the laboratory. In some storehouses, damage by S. paniceum is more serious and establishment of an effective monitoring tool is required. Therefore, in the present study, attractions of these beetles to UV and blue light traps were compared to develop a tool to monitor the beetle pests. First, adult L. serricorne and S. paniceum beetles were provided with UV- and blue-LED panels whose light intensities were equalized in the laboratory, and the walking and flying paths of each adult were recorded and measured. As a result, adults were clearly attracted to the side of UV-LED panel by walking compared to the blue one. Second, we compared the numbers of cigarette beetles collected by sticky sheets that were set in the back of UV or blue-light LED traps in a real storehouse. The results showed that these beetles were significantly more attracted to UV than blue-light LED traps, indicating the UV-LED trap is a powerful tool to monitor these two pest species.

Monitoring and Detecting the Cigarette Beetle (Coleoptera: Anobiidae) Using Ultraviolet (LED) Direct and Reflected Lights and/or Pheromone Traps in a Laboratory and a Storehouse.

The cigarette beetle, Lasioderma serricorne (F.), is an important stored-product pest worldwide because it damages dry foods. Detection and removal of the female L. serricorne will help to facilitate the control of the insect by removal of the egg-laying populations. In this manuscript, we examined the responses by L. serricorne to direct and reflected light in transparent cube (50 m3) set in a chamber (200 m3) and a stored facility with both direct and reflected UV-LED lights. The study also examined the responses by the beetles to light in the presence or absence of pheromone in traps that are placed at different heights. Reflected light attracted more beetles than the direct light in the experimental chamber, but the direct light traps attracted more beetles than the reflected light traps in the storehouse. Pheromone traps attracted only males; UV-LED traps attracted both sexes. The UV-LED traps with a pheromone, i.e., combined trap, attracted more males than UV-LED light traps without a pheromone, whereas the attraction of UV-LED traps with and without the pheromone was similar in females. The results suggest that UV-LED light trap combined with a sex pheromone is the best solution for monitoring and controlling L. serricorne.

Maternal vibration: an important cue for embryo hatching in a subsocial shield bug.

Hatching care has been reported for many taxonomic groups, from invertebrates to vertebrates. The sophisticated care that occurs around hatching time is expected to have an adaptive function supporting the feeble young. However, details of the characteristics of the adaptive function of hatching care remain unclear. This study investigated the hatching care of the subsocial shield bug, Parastrachia japonensis (Heteroptera: Parastrachiidae) to verify its function. Results show that the P. japonensis mothers vibrated the egg mass intermittently while maintaining an egg-guarding posture. Then embryos started to emerge from their shells synchronously. Unlike such behaviors of closely related species, this vibrating behavior was faint, but lasted more than 6 h. To investigate the effect of this behavior on hatching synchrony and hatching success, we observed the hatching pattern and the hatching rate in control, mother-removed, and two artificial vibration groups. Control broods experienced continuous guarding from the mother. Intermittent artificial vibration broods were exposed to vibrations that matched the temporal pattern of maternal vibration produced by a motor. They showed synchronous hatching patterns and high hatching rates. However, for mother-removed broods, which were isolated from the mother, and when we provided continuous artificial vibration that did not match the temporal pattern of the maternal vibration, embryo hatching was not only asynchronous: some embryos failed to emerge from their shells. These results lead us to infer that hatching care in P. japonensis has two functions: hatching regulation and hatching assistance. Nevertheless, several points of observational and circumstantial evidence clearly contraindicate hatching assistance. A reduction in the hatching rate might result from dependence on maternal hatching care as a strong cue in P. japonensis. We conclude that the hatching care of P. japonensis regulates the hatching pattern and serves as an important cue to induce embryo hatching.

Diverse strategies for vertical symbiont transmission among subsocial stinkbugs.

Sociality may affect symbiosis and vice versa. Many plant-sucking stinkbugs harbor mutualistic bacterial symbionts in the midgut. In the superfamily Pentatomoidea, adult females excrete symbiont-containing materials from the anus, which their offspring ingest orally and establish vertical symbiont transmission. In many stinkbug families whose members are mostly non-social, females excrete symbiont-containing materials onto/beside eggs upon oviposition. However, exceptional cases have been reported from two subsocial species representing the closely related families Cydnidae and Parastrachiidae, wherein females remain nearby eggs for maternal care after oviposition, and provide their offspring with symbiont-containing secretions at later stages, either just before or after hatching. These observations suggested that sociality of the host stinkbugs may be correlated with their symbiont transmission strategies. However, we found that cydnid stinkbugs of the genus Adomerus, which are associated with gammaproteobacterial gut symbionts and exhibit elaborate maternal care over their offspring, smear symbiont-containing secretions onto eggs upon oviposition as many non-social stinkbugs do. Surface sterilization of the eggs resulted in aposymbiotic insects of slower growth, smaller size and abnormal body coloration, indicating vertical symbiont transmission via egg surface contamination and presumable beneficial nature of the symbiosis. The Adomerus symbionts exhibited AT-biased nucleotide compositions, accelerated molecular evolutionary rates and reduced genome size, while these degenerative genomic traits were less severe than those in the symbiont of a subsocial parastrachiid. These results suggest that not only sociality but also other ecological and evolutionary aspects of the host stinkbugs, including the host-symbiont co-evolutionary history, may have substantially affected their symbiont transmission strategies.

Species-specific and female host-biased ectophoresy in the roundworm Caenorhabditis japonica.

Caenorhabditis japonica is a bacteriophagous nematode species that was discovered on the semi-social burrower bug, Parastrachia japonensis, which demonstrates egg-guarding and provisioning behaviors. To understand the life history of C. japonica in relation to P. japonensis, we demonstrated the specificity of this association and fluctuations in nematode number on the insect throughout the year. C. japonica dauer larvae (DL), larvae in a nonfeeding diapause stage, were predominantly found as clumps on the adult female insects but rarely found on the male insects in all populations examined. This female-biased association was consistent throughout the year, but after the nymphs hatched, nematodes were not detected on the mother insects showing provisioning behavior. DL appeared on the nymphs, and the number of DL on the newly emerged female insects gradually increased thereafter. C. japonica has never been detected on other invertebrates collected from the P. japonensis habitat thus far. Our data suggest that the life cycles of C. japonica and P. japonensis are synchronized.

Trophic eggs compensate for poor offspring feeding capacity in a subsocial burrower bug.

Various animals produce inviable eggs or egg-like structures called trophic eggs, which are presumed to be an extended maternal investment for the offspring. However, there is little knowledge about the ecological or physiological constraints associated with their evolutionary origin. Trophic eggs of the seminivorous subsocial burrower bug (Canthophorus niveimarginatus) have some unique characteristics. Trophic eggs are obligate for nymphal survival, and first-instar nymphs die without them. To identify the cause of nymphal death, we hypothesized that first-instar nymphs starve to death because they cannot feed on anything but trophic eggs. Although first-instar nymphs fed on artificially exposed endosperm did survive, nymphs that were provided with intact seed were not able to penetrate the seed vessel and starved to death. Another hypothesis that trophic eggs play a role in transferring the midgut symbiont, essential for survival in heteropteran bugs, from mother to offspring was rejected because almost all nymphs had retained the symbiont without feeding on trophic eggs. These results suggest that poor feeding capacity of the offspring is the cause of nymphal death, and the important constraint that promotes the evolution of the curious trophic egg system in C. niveimarginatus.

Phylogenetic position and peculiar genetic traits of a midgut bacterial symbiont of the stinkbug Parastrachia japonensis.

The stinkbug Parastrachia japonensis (Hemiptera: Parastrachiidae) is known for its prolonged prereproductive nonfeeding period, maternal care of eggs in an underground nest, and maternal collection and provisioning of food (fruits) for nymphs. A previous study suggested that a bacterial symbiont is involved in uric acid recycling in this insect during the nonfeeding period, but the identity of this symbiont has not been determined. Here we characterized a novel bacterial symbiont obtained from P. japonensis. Molecular phylogenetic analyses based on 16S rRNA, gyrB, and groEL gene sequences consistently indicated that this symbiont constituted a distinct lineage in the Gammaproteobacteria that has no close relatives but is allied with gut symbionts of acanthosomatid and plataspid stinkbugs, as well as with endocellular symbionts of sharpshooters, tsetse flies, and aphids. The symbiont genes had a remarkably AT-biased nucleotide composition and exhibited significantly accelerated molecular evolution. The symbiont genome was extremely reduced; its size was estimated to be 0.85 Mb. These results suggest that there has been an intimate host-symbiont association over evolutionary time. The symbiont was localized in swollen crypts in a posterior part of the midgut, which was a specialized symbiotic organ. The possibility that the symbiont is involved in uric acid recycling is discussed. The designation "Candidatus Benitsuchiphilus tojoi" is proposed for the symbiont.

Pre- and post-hatch trophic egg production in the subsocial burrower bug, Canthophorus niveimarginatus (Heteroptera: Cydnidae).

In recent years, three terrestrial bugs, Adomerus triguttulus and Sehirus cinctus (Cydnidae) and the closely related Parastrachia japonensis (Parastrachiidae), have been the focus of several fascinating studies because of the remarkable, extensive parental care they were found to display. This care includes egg and nymph guarding, production of trophic eggs, unfertilized, low cost eggs that are used as food by newly hatched nymphs, and progressive provisioning of the host seed. In this study, we have investigated yet a third related Asian cydnid, Canthophorus niveimarginatus, with regard to the possible occurrence of some or all of these complex traits in order to assess how widespread these maternal investment patterns are in this group of insects and to better understand the implications of their manifestations from an evolutionary context. Manipulative experiments were carried out in the lab to determine whether females provision nests. Observational and egg removal studies were carried out to determine whether trophic eggs are produced, and, if they are, their possible impact on nymphal success. The findings revealed that C. niveimarginatus does, in fact, progressively provision young, and this species also displays all of the other behaviors associated with extended parental care in subsocial insects. Moreover, unlike the other two related species, which place trophic eggs on the surface of the original egg mass, C. niveimarginatus produces both pre- and post-hatch trophic eggs. Nymphs deprived of access to post-hatch trophic eggs had significantly lower body weight and survival rate than those that fed on them. To our knowledge, this is the first time the production of both pre- and post-hatch trophic eggs has been demonstrated in insects outside the Hymenoptera. In this paper, we qualitatively and quantitatively demonstrate the provisioning behavior and patterns of trophic egg production in C. niveimarginatus. When and how trophic eggs are produced and delivered to young should have important correlations with the ecological and life history constraints under which a species has evolved. Thus, we also discuss the possible ecological and life history factors that favor the evolution of post-hatch trophic eggs.

Presence of rhodopsin and porphyropsin in the eyes of 164 fishes, representing marine, diadromous, coastal and freshwater species--a qualitative and comparative study.

There are two types of visual pigments in fish eyes; most marine fishes have rhodopsin, while most freshwater fishes have porphyropsin. The biochemical basis for this dichotomy is the nature of the chromophores, retinal (A1) and 3-dehydroretinal (A2), each of which is bound by an opsin. In order to study the regional distribution of these visual pigments, we performed a new survey of the visual pigment chromophores in the eyes of many species of fish. Fish eyes from 164 species were used to examine their chromophores by high-performance liquid chromatography--44 species of freshwater fish, 20 of peripheral freshwater fish (coastal species), 10 of diadromous fish and 90 of seawater fish (marine species) were studied. The eyes of freshwater fish, limb freshwater fish and diadromous fish had both A1 and A2 chromophores, whereas those of marine fish possessed only A1 chromophores. Our results are similar to those of previous studies; however, we made a new finding that fish which live in freshwater possessed A1 if living near the sea and A2 if living far from the sea if they possessed only one type of chromophore.

Canopy compass in nocturnal homing of the subsocial shield bug, Parastrachia japonensis (Heteroptera: Parastrachiidae).

In contrast to an open environment where a specific celestial cue is predominantly used, visual contrast of canopies against the sky through the gap, known as canopy cues, is known to play a major role for visually guided insect navigators in woodland habitats. In this paper, we investigated whether a subsocial shield bug, Parastrachia japonensis, could gauge direction using canopy cues on a moonless night. The results show that they could perform the round trip foraging behaviour even in an experimental arena with only an artificial round gap opened in the ceiling of the arena and adjust their homing direction for a new azimuth when the gap was rotated. Thus, P. japonensis can use slightly brighter canopy cues as a compass reference but not complex landmarks during nocturnal homing behaviour.

Water uptake via two pairs of specialized legs in Ligia exotica (Crustacea, Isopoda).

When individuals of Ligia exotica were exposed to dry environments, their weight decreased within 3 h to 90% of the initial weight. When the animals were subsequently presented with wet paper, pereiopods VI and VII were firmly apposed and moved around until their tips were stationary for some seconds. Subsequently the pleopods became soaked in water and the body weight recovered. Morphological observations revealed hollow structures on the surface of the dactylus and propodus of pereiopods VI and VII, and a series of thin cuticular protrusions, oriented in several parallel lines, from the propodite to the ischiopodite of pereiopod VI and on the basipodite of pereiopod VII. The width between the parallel lines varied little, but the total width of the regular lines increased linearly with increasing body size. When isolated caudal pereiopods were immersed, beginning at the distal end, in water, water flowed along pereiopod VI as far as the proximal end of the ischiopodite, but water flow along pereiopod VII occurred only in conjunction with the basipodite. This means that water uptake with the legs requires both pereiopods VI and VII and can be achieved only when these legs are closely apposed.

Round-the-clock homing behavior of a subsocial shield bug, Parastrachia japonensis (Heteroptera: Parastrachiidae), using path integration.

Females of the subsocial shield bug, Parastrachia japonensis (Parastrachiidae), are central-place foragers, collecting drupes for their young from nearby host trees by walking along the forest floor both during the day and at night. Because burrows are often some distance from the drupe-shedding tree, the bugs must repeatedly leave their burrows, search for drupes, and return to the burrows. After a bug leaves its burrow, it searches arduously until it encounters a drupe. When a drupe is obtained, the bug always takes the shortest route back to its burrow. It has been clarified that this bug utilizes path integration during diurnal provisioning excursions. In this paper, we examined nocturnal behavior and some parameters of the path integration utilized by P. japonensis. There were no observable differences between day and night in the patterns of foraging and direct-homing behavior. When the bug was displaced to another position during the day or night, it always walked straight toward the fictive burrow, the site where the burrow should be if it had been displaced along with the bug, and then displayed searching behavior in the vicinity of the fictive burrow. The distance of the straight run corresponded accurately with a straight line between the burrow and the place where the bug obtained the drupe. These results indicate that P. japonensis orients toward the burrow using path integration both during diurnal and nocturnal provisioning behavior.

The directional homing behaviour of the subsocial shield bug, Parastrachia japonensis (Heteroptera: Cydnidae), under different photic conditions.

The female subsocial shield bug, Parastrachia japonensis, provisions its nymphs by foraging on the ground in the forest during the Japanese rainy season, and the bug uses homing navigation to drag a drupe back to its burrow by the shortest route during the day. To study whether or not this bug performs this provisioning behaviour under different photic conditions, we observed the homing behaviour and homing direction of bugs in the field around the clock and/or under various weather conditions. The bugs foraged the whole day during the busiest provisioning period, and the number of walking bugs was not affected by the different weather conditions. Such navigational behaviour, regardless of the time of the day and the weather conditions, is rare in insect navigation. To test whether the bug uses visual cues, we covered the compound eyes and ocelli with opaque or clear paint just before homing began. During the day and at night, and in all weather conditions, the homing direction of blind bugs, but not those with clear-painted eyes was disoriented, indicating that this species uses visual cues dominantly under all photic conditions.